Expression of immune response genes in rainbow trout skin induced by Gyrodactylus derjavini infections

By means of semi-quantitative RT-PCR, expression of a number of immune relevant genes was studied in skin of small rainbow trout Oncorhynchus mykiss (Walbaum, 1792) fry during both primary and secondary infections with the ectoparasitic monogenean Gyrodactylus derjavini Mikailov, 1975. The target genes studied included the cyto- and chemokines TNF-alpha1, TNF-alpha2, TGF-beta and IL-8, the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) genes and finally, two cell markers, the beta-chains of TCR and MHC II, from the adaptive arm of the immune system. In general, constitutive expression of all studied genes was apparent. Significant increases in expression of the TNF-alpha1 isoform could be observed at day 8 p.i. in primary infections and although less marked, the alpha2 isoform, of TNF showed a similar trend. With the cytokine TGF-beta, 8-10 times increase in the transcription levels was observed in secondary infections compared to uninfected hosts. However, no parasite related changes in expression patterns could be observed for IL-8. Parasite infections elicited strong iNOS expression by 4 days p.i., but significant differences were not detected before day 8 p.i., when transcript levels were increased 5.5-9.6 times compared to uninfected controls. Augmented expression of COX-2 could also be observed in primary, but not secondary, infections at later stages of infections. No clear parasite related changes in transcript levels of the two cell markers TCRbeta and MHC IIbeta could be observed. Although the cellular source(s) was not determined, most of the examined factors appear to take part in a local signalling network of pivotal importance for the initiation, orchestration, effectuation and modulation of immune responses in rainbow trout against the ectoparasite G. derjavini. (C) 2003 Elsevier B.V. All rights reserved.